Abstract

Introduction: Biomedical applications of nanoparticles (NPs) as enzyme inhibitors have recently come to light. Oxides of metals native to the physiological environment (eg, Fe, Zn, Mg, etc.) are of particular interest—especially the functional consequences of their enzyme interaction.

Materials and methods: Here, Fe 2 O 3 , zinc oxide (ZnO), magnesium oxide (MgO) and nickel oxide (NiO) NPs are compared to copper (Cu) and boron carbide (B 4 C) NPs. The functional impact of NP interaction to the model enzyme luciferase is determined by 2-dimensional fluorescence difference spectroscopy (2-D FDS) and 2-dimensional photoluminescence difference spectroscopy (2-D PLDS). By 2-D FDS analysis, the change in maximal intensity and in 2-D FDS area under the curve (AUC) is in the order Cu~B 4 C>ZnO>NiO>>Fe 2 O 3 >MgO. The induced changes in protein conformation are confirmed by tryptic digests and gel electrophoresis.

Results: Analysis of possible trypsin cleavage sites suggest that cleavage mostly occurs in the range of residues 112–155 and 372–439, giving a major 45 kDa band. By 2-D PLDS, it is found that B 4 C NPs completely ablate bioluminescence, while Cu and Fe 2 O 3 NPs yield a unique bimodal negative decay rate, −7.67×10 3 and −3.50×10 1 relative light units respectively. Cu NPs, in particular, give a remarkable 271% change in enzyme activity. Molecular dynamics simulations in water predicted that the surfaces of metal oxide NPs become capped with metal hydroxide groups under physiological conditions, while the surface of B 4 C becomes populated with boronic acid or borinic acid groups. These predictions are supported by the experimentally determined zeta potential. Thin layer chromatography patterns further support this conception of the NP surfaces, where stabilizing interactions were in the order ionic>polar>non-polar for the series tested.

Conclusion: Overall the results suggest that B 4 C and Cu NP functional dynamics on enzyme biochemistry are unique and should be examined further for potential ramifications on other model, physiological or disease-relevant enzymes.

Document Type

Article

DOI

https://doi.org/10.2147/IJN.S152222

Rights Information

© 2018 The authors. This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.

Keywords

2-D FDS, 2-dimensional fluorescence difference, AUC, Corrected light intensity emitted, Emission wavelength, Excitation wavelength, RLU, Wavelength of max intensity

Publication Date

1-1-2018

Journal Title

International Journal of Nanomedicine

Share

COinS